Western Blot - Theory and method

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  • Опубликовано: 4 ноя 2024

Комментарии • 52

  • @Vekey2
    @Vekey2 5 лет назад +6

    This video was everything I wanted! I have a vague idea of everything in here and just needed a refresher on the actual practical use of everything. Thank you!

  • @SerenSanguis
    @SerenSanguis 3 года назад +20

    Very nice! The closest thing we have to lab practice in the pandemic!

  • @vishk123
    @vishk123 2 года назад

    Thank you for inviting me to your game! I played and learnt a lot!

  • @hirosewill893
    @hirosewill893 7 лет назад +4

    Nice protocol and instruction. Thank you for the video

  • @amymay16
    @amymay16 2 года назад +1

    Thanks, great video! How does the metal stirrer help the process? I usually leave the tank without it being stirred the whole time, would love to know how stirring it helps as I may try that! Thanks so much in advance.

  • @prembobby112
    @prembobby112 2 года назад

    Everything clearly explained to more useful to practice this Western Blotting
    Thanking you Sir/Mam

  • @DocScoop7
    @DocScoop7 11 месяцев назад

    it was such an awesome video

  • @RoyRahebKhelo
    @RoyRahebKhelo 6 лет назад +3

    Splendid presentation, thank you.

  • @tle6974
    @tle6974 7 лет назад +33

    Good video but please
    14:51 dont hold the pipette upside down with a used tip on it... ;)

    • @travstravinho2766
      @travstravinho2766 7 лет назад +1

      Yas omg

    • @JaLLaM86
      @JaLLaM86 6 лет назад +1

      ... Or use a computer mouse with gloves on.

    • @tylerlord1340
      @tylerlord1340 6 лет назад +2

      can you explain why?

    • @bradywells1293
      @bradywells1293 6 лет назад +4

      The liquid can make its way into the pipette and cause contamination to the next sample(s) you use it for or it can just damage the sensitivity of the pipette by getting stuff gunked up inside.

    • @KalahSimpson
      @KalahSimpson 6 лет назад +2

      Better yet, use filter tips

  • @ماجدفرج-خ4ط
    @ماجدفرج-خ4ط 5 месяцев назад

    Nice video, how long for blot
    ting at 115 volt?

  • @Lina-7578
    @Lina-7578 6 лет назад +2

    Thank you! Great video, it helps me a lot!

  • @jfromtheusa
    @jfromtheusa Год назад

    hugely helpful

  • @dishabanerjee9630
    @dishabanerjee9630 8 месяцев назад

    Thank you so much!!

  • @Biomeducated
    @Biomeducated 5 лет назад +6

    Great tutorial! But,
    13:58 you didn't depict the amplification tree (usually the secondary is polyclonal), so there's multiple secondaries binding to the primary to 'blow up' your signal from the HRP reaction.
    Besides that: *great channel* here, man! Why did you stop though :(
    Just as I'm discovering all these helpful channels... I even made a Top 10 channel overview on my own channel, and I'm planning on making a second one with 'smaller channels' like this one. All good stuff for Biomedical/Life Science students!

    • @lukehebert6207
      @lukehebert6207 3 года назад +1

      I'm re-acquainting myself with this stuff after years of not being in a wet lab. Thanks for this detail and the overview vid on your channel. Polyclonal secondary antibodies sounds clever and makes a lot of sense.

    • @Biomeducated
      @Biomeducated 3 года назад

      @@lukehebert6207 Cheers, man!

  •  5 лет назад

    Very useful video. Thank you.

  • @BalajiHariPhD
    @BalajiHariPhD 2 месяца назад

    I'm interested to do this western blot techniques

  • @digitalmediazone9717
    @digitalmediazone9717 Год назад

    Thank you

  • @valentinascatizzi6406
    @valentinascatizzi6406 4 года назад

    thank you, brilliant and accurate explanation

  • @汪华志
    @汪华志 6 лет назад +6

    i have a question : how tween inhibits detection ? what is the reaction mechanism

    • @imranchandler1083
      @imranchandler1083 3 года назад

      you prolly dont give a damn but does someone know of a way to get back into an Instagram account?
      I was stupid lost my account password. I would appreciate any tips you can offer me!

    • @axtonjax3700
      @axtonjax3700 3 года назад

      @Imran Chandler Instablaster ;)

    • @imranchandler1083
      @imranchandler1083 3 года назад

      @Axton Jax thanks for your reply. I found the site thru google and im trying it out atm.
      I see it takes quite some time so I will reply here later with my results.

    • @imranchandler1083
      @imranchandler1083 3 года назад

      @Axton Jax it did the trick and I now got access to my account again. Im so happy:D
      Thanks so much, you saved my account !

    • @axtonjax3700
      @axtonjax3700 3 года назад

      @Imran Chandler You are welcome xD

  • @ChristyLuvz
    @ChristyLuvz 6 лет назад +1

    Can the primary and secondary antibodies be diluted in the 5% non-fat milk and then each incubated with the membrane? Rather than doing the blocking step as its on whole step?

    • @Biomeducated
      @Biomeducated 5 лет назад +1

      You mean put everything in at once at the same time to reduce protocol time? Not with the blocking step for sure, but perhaps you can premix primary and secondary at equimolar concentrations, then you are sure that no excess secondary will deplete your primary if you wash afterwards. Then again, you would loose your 'amplification tree' which could results in not seeing a good signal. Usually the secondary antibody is a polyclonal so multiple HRP-conjugated antibodies bind to the Fc-part of your primary, needed to amplify your signal. I would just stay with the sequential protocol.
      Or maybe I misread your question (just realized I think): yes, your antibodies should be diluted in the same solution as the blocking, but usually it's more diluted than for the blocking. So if you block with 5%, you stain in 1% or 0.5%. Note that some protocols recommend using BSA as blocking and for dilution...

    • @МаринаЗеленюк-р3ч
      @МаринаЗеленюк-р3ч 6 месяцев назад

      Usually you should to incubate the membrane with bloaking solution during 1 hour. Then you should wash 3 times and incubate with primary Ab diluted in 5% milk PBS overnight. Then you schould wash 3 times and incubate with secondary Ab diluted in 5% milk PBS during 1 hour. Wash 3 times and make a picture.

  • @Kysersozé1
    @Kysersozé1 3 года назад

    Thank you.

  • @SixTough
    @SixTough 7 лет назад +1

    excellent

  • @rishavdas9939
    @rishavdas9939 7 лет назад +2

    Good

  • @nevinvolminck7920
    @nevinvolminck7920 3 месяца назад

    my mebrane disloved immedietly in the methanol :(

  • @kursatrecepdeniz705
    @kursatrecepdeniz705 5 месяцев назад +1

    Balls🙏🏻

  • @detroit7543
    @detroit7543 6 лет назад +3

    I wish you have subtitle..

  • @Buongona
    @Buongona 6 лет назад +3

    so blotting is blotting? I guess I really need to dig deeper...

    • @selamioguz6111
      @selamioguz6111 4 года назад

      Ok I'll see what the weather was not in my head is

  • @powerharriet
    @powerharriet 3 года назад

    Hair down in the lab?

  • @Airisweetheart
    @Airisweetheart 6 лет назад +11

    Dont they make you put your hair up in a lab?

    • @richardwu9013
      @richardwu9013 6 лет назад +1

      Airisweetheart only with caustic or dangerous substances.

    • @SerenSanguis
      @SerenSanguis 3 года назад +2

      Well, personally I always do wether I am asked to or not, because they get in the way and I can't concentrate

  • @lincolnoliveira3041
    @lincolnoliveira3041 3 года назад

    Lcnd